Bacteria synthesize a plethora of complex surface-associated polysaccharides which enable them to persist and thrive in distinct niches. These glycans serve an array of purposes pertaining to virulence, colonization, antimicrobial resistance, stealth, and biofilm formation. The Wzx/Wzy-dependent pathway is universally the predominant system for bacterial polysaccharide synthesis. This system is responsible for the production of lipopolysaccharide (LPS) O-antigen (Oag), enterobacterial common antigen, capsule, and exopolysaccharides, with orthologs present in both Gram-negative and Gram-positive microbes. Studies focusing principally on Pseudomonas, Shigella, and Salmonella LPS Oag synthesis have provided much of the framework underpinning the biochemical and molecular mechanism behind polysaccharide synthesis via this pathway. LPS Oag production via the Wzx/Wzy-dependent pathway occurs through the stepwise activity of multiple key biosynthetic enzymes, including primarily the polymerase, Wzy, which is responsible for the Oag assembly, and the polysaccharide co-polymerase, Wzz, which effectively modulates the length of the glycan produced. In this review, we provide a comprehensive summary of the latest genetic, structural, and mechanistic data for the main protein candidates of the Wzx/Wzy-dependent pathway, in addition to an examination of their substrate specificities. Furthermore, we have reviewed recent insights pertaining to the dynamics/kinetics of glycan synthesis by this mechanism, including the interplay of the key proteins among themselves and in complex with their substrate. Lastly, we outline key gaps in the literature and suggest future research avenues, with the aim to stimulate ongoing research into this critical pathway responsible for the production of key virulence factors for numerous debilitating and lethal pathogens.